TRPV4在左炔诺孕酮致输卵管妊娠发病中的作用及其机制

Ectopic pregnancy is a kind of common acute abdomen which endangers women's life. In our previous studies, we found that levonorgestrel (LNG) increased the risk of tubal pregnancy, however the mechanism is unclear. LNG can reduce the tubal ciliary beat frequency and the expression of TRPV4 in tubal epithelium. In tubal epithelium from women diagnosed with tubal pregnancy, the expression of TRPV4 was reduced and was inversely related to progesterone level in serum. These results suggested that TRPV4 may be involved in the occurrence of tubal pregnancy. Starting with TRPV4, we intend to clarify the correlation between TRPV4 and LNG and tubal pregnancy by using molecular biotechnology; We plan to investigate the effect of TRPV4 on tubal epithelial structure and function and tubal ciliary beat frequency in primary cultured ciliated cells, using electron microscope, tubal ciliary testing platform and cellular electrophysiology; We are going to study the molecular mechanism of down-regulation of TRPV4 by LNG in cell models by applying chromatin immunoprecipitation, luciferase reporter gene and protein profiling; In order to provide theoretical basis concerning on the pathogenesis of tubal pregnancy following contraception failure, we plan to apply CRISPER/Cas9 to construct a TRPV4 conditional-knockout mouse model and use living radiological technology to study the effect of LNG, TRPV4 and intracellular calcium signaling pathways on tubal ciliary movement and embryo transportation.

异位妊娠是危及妇女生命的常见急腹症，危害大，我们既往研究发现左炔诺孕酮（LNG）增加异位妊娠风险，其机制尚不清楚。前期研究显示LNG可引起输卵管纤毛运动下降，且可降调人输卵管上皮TRPV4表达；TRPV4在输卵管妊娠中表达下调，且与血清孕酮水平负相关。提示TRPV4可能参与LNG所致输卵管妊娠发生。本项目以TRPV4为切入点，利用分子生物学方法明确TRPV4与LNG及输卵管妊娠间关联；利用纤毛检测平台及电生理等技术在原代纤毛细胞上研究TRPV4对输卵管上皮细胞形态、功能和纤毛摆动频率的影响；利用染色质免疫共沉淀、荧光素酶报告基因、蛋白质谱等技术在细胞模型上研究LNG下调TRPV4的分子机制；CRISPR/Cas9分别构建条件性敲除TRPV4小鼠模型，应用活体影像技术明确LNG/TRPV4/胞内钙信号通路对输卵管纤毛运动及胚胎运输的影响，为LNG所致输卵管妊娠的发生机制提供理论依据。

异位妊娠是危及妇女生命的常见急腹症，危害大，我们既往研究发现左炔诺孕酮（LNG）增加异位妊娠风险，其机制尚不清楚。前期研究显示LNG可引起输卵管纤毛运动下降，且可降低人输卵管上皮TRPV4表达；TRPV4在输卵管妊娠中表达下调，且与血清孕酮水平负相关，提示TRPV4可能参与LNG所致输卵管妊娠发生。本项目研究发现LNG通过抑制TRPV4及其介导的细胞钙离子内流降低纤毛摆动频率，引起胚胎滞留，导致输卵管妊娠的发生，揭示了LNG诱发输卵管妊娠发生的分子机制，为临床降低LNG诱导的EP风险提供新线索。